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2. Materials And Methods 2.1. Chemicals And Drugs Acaie Extract Was Purchased From Holland & Barrett / Uk And Other Chemical Reagents Were Purcha

792 words - 4 pages

2. Materials and Methods

2.1. Chemicals and drugs
Acaie extract was purchased from Holland & Barrett / UK and other chemical reagents were purchased from sigma Aldrich chemical co. (st. Louis, MO., USA).

2.2. Animals

Forty male Sprague-Dawley rats weighing 180–200 g were housed at cages in a temperature-controlled (25±1ºC) environment and provided free access to pelleted food and purified drinking water ad libitum, and left to accommodate for one week before the experiment. The animal experiments described later were approved by the Ethics Committee, Faculty of Pharmacy, Helwan University, Cairo, Egypt.
2.3. Experimental design
The animals were divided into 4 groups, ten per each group, ...view middle of the document...

During ischemia and reperfusion, the wound was temporarily closed to maintain body temperature. Occlusion was verified visually by change in the color of the kidneys to a paler shade and once the reperfusion began, the renal color turned to red. Other rats were subjected to sham operation, undergo identical surgical operation, except for clamping and were maintained under anaesthesia for the duration of the experiment (45min +4 h). At the end of all experiments, rats were disposed of by a sodium pentobarbital overdose. Blood samples were collected immediately from the aorta; the plasma was separated by centrifugation at 3000 rpm for 15 min at 4°C. These samples were used for measurements of renal functional parameters. Also the kidneys were removed and decapsulated; plasma and kidney samples were frozen at -70°C until analysis.

2.4. Renal functional parameters
2.5. Estimation of lipid peroxidation—malondialdehyde (MDA)
Renal malondialdehyde (MDA) levels (an end product of lipid peroxidation), a specificmarker for the oxidative damage in proteins were estimated accordingly to (Mihara and Uchiyama, 1978).

2.6. Measurement of cytokines levels
Cytokines levels were determined by ELISA kits specific for rats. The parameters evaluated were INF-γ, IL-10. Each sample was assayed in duplicate. The concentrations of cytokines were determined with the help of a standard curve.
INF-γ
The activity of kidney caspase-3 was measured by using a colorimetric assay kit (Uscn Life Science...

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